Aspergillus flavus strains and mixed flora that do not produce aflatoxin and their application

A kind of technology of aflatoxin and Aspergillus flavus strains, applied in the direction of application, fungi, and methods based on microorganisms, etc., can solve the problems of unstable control effect, reduce unstable control effect, expand the scope of applicable regions, and improve utilization rate Effect

Inactive Publication Date: 2017-10-31
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The application of a single biocontrol bacteria in the field often results in unstable control effects due to the fitness of the biocontrol strains

Method used

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  • Aspergillus flavus strains and mixed flora that do not produce aflatoxin and their application
  • Aspergillus flavus strains and mixed flora that do not produce aflatoxin and their application
  • Aspergillus flavus strains and mixed flora that do not produce aflatoxin and their application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 does not produce the isolation of the aspergillus flavus strain of aflatoxin, identification

[0032] 1. Isolation of Aspergillus flavus strains from soil, corn, peanut and other habitats

[0033] Soil was collected from corn and peanut fields in Jiangsu, Zhejiang, Shandong, Fujian, Anhui and other places, and the soil was cultured with YES medium to isolate Aspergillus flavus strains. Ingredients per liter of YES medium: Yeast extract 1g, sucrose 10g, NaCl 60g, agar 20g, add CuSO after sterilization 4 .ZnSO 4 1 mL, 0.4% clonamide 5 ml, streptomycin 100 μg / mL.

[0034] 2. Use bioassay, thin-layer chromatography and enzyme-linked immunoassay to analyze the aflatoxin production of the strain.

[0035] details as follows:

[0036] Bioassay method: Inoculate the isolated Aspergillus flavus strain on YES medium containing 0.3% cyclodextrin, culture it at 30°C for 7 days, fumigate the colony with 25% ammonia water for 3 minutes, if the back of the colony is pin...

Embodiment 2

[0053] Identification of aflatoxin synthetic gene deletion in the aspergillus flavus strains A 051, A F 052 and A F 053 that do not produce aflatoxin in embodiment 2

[0054] (1) Primer synthesis

[0055] In order to clarify the deletion of the aflatoxin synthesis gene in the non-aflatoxin-producing Aspergillus flavus strains A051, AF052 and AF053, the test refers to the literature Perng-Kuang Chang (ChangP.K., Horn, B.W.and Dorner, J.W. (2005) Sequence Breakpoints in the aflatoxin biosynthesis gene cluster and flanking regions in nonaflatoxigenic Aspergillus flavus isolates. Fungal GenetBio 42, 914-923.) Design primers for the identification of toxin synthesis genes, and identify the deletion of related genes in strains A051, AF052 and AF053 .

[0056] (2) Gene deletion identification

[0057] Using the extracted DNA of Aspergillus flavus strains A051, AF052 and AF053 as templates, conventional PCR reactions were carried out with the above primers, and each reaction had a D...

Embodiment 3

[0061] Example 3 Indoor test evaluation of the inhibitory effect of the Aspergillus flavus biocontrol strain group that does not produce aflatoxin on the Aspergillus flavus producing aflatoxin on corn

[0062] (1) Preparation of corn

[0063] Weigh 20 g of corn, crush it with a juice extractor, and sterilize at 121° C. for 30 minutes before use.

[0064] (2) Preparation and cultivation of bacterial suspension

[0065] The non-toxin-producing Aspergillus flavus strains A051, AF052, AF053 and the aflatoxin-producing Aspergillus flavus standard strain NRRL3357 were respectively inoculated on PDA medium (200 g of potatoes, 20 g of glucose, 20 g of agar, and adding water to 1 L), and the plate was placed at 30 After culturing at ℃ for 5 days, add 0.1% Tween 20 to wash and collect spores, use a vortex shaker to shake and mix, and then adjust the spore concentration with a hemocytometer. A total of 5 treatments were set up in the experiment, as follows:

[0066]Treatment 1: The sp...

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Abstract

The invention discloses a non-aflatoxin-producing Aspergillus flavus strain and mixed flora and application thereof. The strains are Aspergillus flavus AF052, the preservation number is CGMCC NO.9858, and the preservation date is November 20, 2014 , and Aspergillus flavus AF053, deposit number CGMCC NO.9859, deposit date November 20, 2014. Both the strain and the flora can effectively inhibit the formation of aflatoxin in corn and peanuts and improve the quality of agricultural products; in addition, A051, AF052 and AF053 can also secrete cellulase, and the corn treated with the biocontrol flora prepared by using the Aspergillus flavus When the straw and straw are used as animal feed, the biocontrol bacteria carried will produce cellulase, which can degrade cellulose and improve the utilization efficiency of animal feed.

Description

technical field [0001] The invention relates to the field of agricultural product safety, in particular to a non-aflatoxin-producing Aspergillus flavus strain, a mixed flora and an application thereof. Background technique [0002] Aflatoxins are mainly metabolites of Aspergillus flavus and Aspergillus parasiticus, and are the most toxic type of biotoxins found so far in contaminated agricultural products. Among them, aflatoxin B1 is the most toxic and harmful, and its toxicity is 10 times that of potassium cyanide and 68 times that of arsenic. It is listed as a highly toxic substance under strict control. Aflatoxin contamination of agricultural products has become an important pollutant affecting food safety and endangering people's health. [0003] According to the World Food and Agriculture Organization estimates, currently 25% of the world's crop products are contaminated with mycotoxins, mainly aflatoxins. In recent years, countries around the world have not only form...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14A23L3/3571C12R1/67
CPCC12N1/145C12R2001/67
Inventor 陈云尹燕妮马忠华
Owner ZHEJIANG UNIV
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